PhD Scientific Days 2025

Molecular Medicine IV.

Diameter, protein-to-lipid ratio, and lipid signature as defining parameters of Extracellular Vesicles and Non-Vesicular Extracellular Particles

Name of the presenter

Ghosal Sayam

Institute/workplace of the presenter

Department of Genetics, Cell, and Immunobiology

Authors

Sayam Ghosal¹

1: Department of Genetics, Cell and Immunobiology, Semmelweis University

Text of the abstract

Introduction: Extracellular vesicles (EVs) play key roles in intercellular communication and disease progression. However, distinguishing their origins, biogenesis, and functions, along with those of non-vesicular extracellular particles (NVEPs), remains a major challenge.
Aim: This study aims to systematically characterise the origins, structure, and markers of distinct EV subtypes and NVEPs to clarify their release, diversity, and functions.
Methods: We conducted a meta-analysis of ILVs, EVs, and NVEPs using TEM, NTA, SP-IRIS, MACSPLEX, Raman spectroscopy, biochemical assays, lipidomics, high-resolution flow cytometry, and advanced microscopy. These methods allowed us to compare their physical and biochemical properties across wild-type and marker-expressing cells.
Results: Our analyses reveal that large EVs of more than 200nm diameter may not be generated via the exosomal pathway, as ILVs had a mean diameter of 109.60 nm and a maximum diameter of 197.00 nm. NVEPs, exhibited distinct biochemical and lipidomic profiles, with significantly higher protein and lipid content than classical EVs. Furthermore, conventional EV markers were found to be differentially distributed across subpopulations and were not exclusive to any single class or subcellular origin. These findings highlight the considerable heterogeneity among extracellular particles and challenge current classification standards.
Conclusion: Our study advances the understanding of the diversity and complexity of extracellular particles. The results underscore the necessity for refined classification criteria and more precise molecular markers to distinguish between EV subtypes and NVEPs. This work provides a foundation for future research to elucidate the functional roles of these particles in health and disease.
Funding: The project has received funding from the EU’s Horizon 2020 Research and Innovation Programme under Grant agreement No. 739593, OTKA FK 147023, EXCELLENCE 151417, 2019-2.1.7-ERA-NET-2021-00015, and NVKP_16-1-2016-0004 grants of the Hungarian National Research, Development and Innovation Office (NKFIH), the Higher Education Excellence Program (FIKP) and the Therapeutic Thematic Programme TKP2021-EGA-23, RRF-2.3.121-2022-00003 (National Cardiovascular Laboratory Program), VEKOP-2.3.2-162016-00002, VEKOP-2.3.3-15-2017-00016, EKÖP-2024-237 and Stipendium Hungaricum Scholarship 2021.